Substituted 7-acetylamino cephalosporanic acids



United States Patent r 3,345,367 SUBSTITUTED 7-ACETYLAMINOQEPHALOSPORANIC ACIDS Martin Leon Sassiver, Pearl River, and BenjaminArthur Lewis, Suifern, N.Y., and Robert Gordon Shepherd, Ridgewood, NJ,assignors to American Cyanamid Company, Stamford, Conn., a corporationof Maine No Drawing. Filed Feb. 15, 1967, Ser. No. 616,176 10 Claims.(Cl. 260-243) ABSTRACT OF THE DISCLOSURE This disclosure describescompounds of the class of 7 (5,6,7,8 tetrahydronaphthyl 1oxyacetylarnino) cephalosporanic acids and7-(5,6,7,8-tetrahydronaphthyl- 2-oxyacetyla-mino)cephalosporanic acids,useful as antibacterial agents.

Brief summary of the invention This invention relates to new derivativesof 7-aminocephalosporanic acid and, more particularly, is concerned withnovel compounds which may be represented by the following generalformulae:

wherein R R R R R and R are each selected from the group consisting ofhydrogen, methyl, chloro and bromo; A is acetoxy or N-pyridinium; and Mis hydrogen, pharmaceutically acceptable non-toxic cations or an anioniccharge when A is N-pyridinium.

Detailed description of the invention In the general Formulae (I) and(11) set forth above, in those instances where A is N-pyridinium, thecationic charge on this group is matched by the anionic charge of thecarboxylic acid radical, the entire molecule being of a zwitterionnature and M is thus an anionic charge. Typical5,6,7,8-tetrahydronaphthyloxyacetyl groups contemplated by the presentinvention are, for example, 5,6,7,8-tetrahydronaphthyl-l-oxyacetyl,2-chloro-S,6,7,8- tetrahydronaphthyl-l-oxyacetyl,3-bromo-5,6,7,8-tetrahydronaphthyl-l-oxyacetyl,4-bromo-5,6,7,8-tetrahydronaphthyl-1-oxyacetyl,2,4-dichloro-5,6,7,8-tetrahydronaphthyl- 1 oxyacetyl, 2 methyl 5,6,7,8tetrahydronaphthyl 1- oxyacetyl,3,4-dimethyl-5,6,7,8-tetrahydronaphthyl-l-oxyacetyl, 1 methyl 5,6,7,8tetrahydronaphthyl 2 oxyacetyl, 3,4 dimethyl 5,6,7,S tetrahydronaphthyl2- oxyacetyl, 5,6,7,8-tetrahydronaphthyl-2-oxyacetyl, l-chlo r0 5,6,7,8tetrahy-dronaphthyl 2 oxyacetyl, 3 chloro-5,6,7,8-tetrahydronaphthyl-Z-oxyacetyl, 4-bromo-5,6,7,8tetra'hydronaphthyl-Z-oxyacetyl,1,3-dichloro-5,6,7,8-tetrahydronaphthyl-Z-oxyacetyl, and the like.

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Also embraced within the scope of the present invention are thenon-toxic pharmaceutically acceptable salts of these derivatives of7-aminocephalo-sporanic acid. The cations comprised in these salts andembraced by M include, for example, the non-toxic metal cations such asthe sodium ion, potassium ion, calcium ion, magnesium ion as well as theorganic amine cations, such as the tri(lower alkyl)amine cations (e.g.triethylamine), procaine, and the like;

The novel compounds of the present invention, when A is acetoxy inFormulae (I) and (II) above, may be readily prepared by acylating7-aminocephalo'sporanic acid with a compound which may be represented bythe following general formulae:

wherein R R R R R and R are as hereinbefo-re defined and Z is a halide(preferably chloride), azide, acyloxy or p-nitrophenoxy group. Thisacylation of the 7-aminocephalosporanic acid is performed, for example,by the Schotten-Baumann method, taking into consideration thesensitivity of these compounds, under mild conditions and advantageouslyin the presence of a diluent or solvent such as Water or an organicsolvent, for example, a ketone such as acetone, an ether such astetrahydrofuran, or a halogenated hydrocarbon such as chloroform ormethylene chloride. The reaction is preferably conducted in the presenceof a basic agent such as sodium bicarbonate or potassium bicarbonate, oran organic base such as one of the organic bases listed hereinbefore.The reaction is also preferably carried out at a temperature of fromabout 0C. to about 25 C., preferably at 0 C.-

5 C., and over a period of time of a few hours or more.

The acylating agents corresponding to the formulae (III) and (IV), whennew, may be prepared by methods well-known in the art from thecorresponding acids (Z is hydroxy). Thus, the acid can be treated withthionyl chloride or oxalyl chloride, if desired in the presence ofdimethylformamide, to yield the corresponding acyl chlorides (Z ischlorine), which, if desired, can be converted to the acyl azides (Z isN by treatment with sodium azide. The p-nitrophenyl esters (Z isp-nitrophenoxy) can be prepared by following the procedure of Bodanszkyet al. (Biochemical Preparations, vol. 9, p. 110, 1962, John Wiley andSons, New York, N.Y.). Specific acylating agents operable in thisprocess include, for example, 5,6,7,S-tetrahydronaphthyl-l-oxyacetylchloride, 2-chloro- 5,6,7,S-tetrahyd-ronaphthyl-l-oxyacetyl bromide,3-methyl-S,6,7,8-tetrahydronaphthyl-l-oxyacetyl azide, p-nitrophenyl4-bromo-5,6,7,S-tetrahydronaphthyl-l-oxyacetate, 2,4-dichloro-5,6,7,8tetrahydronaphthyl 1 oxyacetyl chloride, 5,6,7,8tetrahydronaphthyl-Z-oxyacetyl chloride, 1 chloro5,6,7,8-tetrahydronaphthyl-Z-oxyacetyl bromide, 3 methyl5,6,7,8-tetrahydronaphthyl-2-oxyprepared from the correspondingsubstituted tetrahydronaphthols by treatment with chloroacetic acid.

The novel compounds of the present invention, when A is N-pyridinium inFormulae (I) and (II) above, may be readily prepared from thecorresponding 7-aminocephalosporanic acid derivatives (A is acetoxy inFormulae (I) and (11) above) by treatment with pyridine in water oraqueous acetone at 2050 C. and for a period of time of about 13 days.The resulting 3-(1-pyridylmethyl)-3-cephem-4-carboxylic acid betainesmay then be isolated by standard procedures of precipitation andcrystallization.

Depending on the reaction conditions used, the new compounds of thepresent invention are obtained in the free form or in the form of theirsalts. From the salts it is possible to prepare the acids in knownmanner, or from the acids the salts are readily accessible, for example,by reaction with hydroxides, carbonates or bicarbonates of alkali metalsor alkaline earth metals, or with organic amines.

The novel compounds of the present invention are biologically active andhave been found to possess antibacterial activity. As indicated, theyare useful antimicrobial agents and have high antimicrobal activity invitro against standard laboratory microorganisms used to screen foractivity against pathogens. The antibacterial spectrum of typicalcompounds of the present invention, representing the concentrationrequired to inhibit the growth of various typical bacteria, wasdetermined in a standard manner by the agar-dilution streak-platetechnique which is commonly used in testing new antibiotics. Thefollowing table summarizes the in vitro activity of 7 (5,6,7,8tetrahydronaphthyl 1 oxyacetylamino) cephalosporanic acid (1),7-(5,6,7,S-tetrahydron-aphthyl- 2-oxyacetylamino)cephalosporanic acid(2), and 7-(2,4- dichloro-5,6,7,8-tetrahydronaphthyl l-oxyacetylamino)cephalosporanic acid (3) as compared with Cephalosporin C (4) against avariety of disease-causing microorganisms.

Minimal inhibitory cone. (meg. m1.)

Organisms Staphylococcus aureus No. 11 0.39 0.2 0.2 100 Staphylococcusaurcas ATCC 13709 0.1 0. 05 0. 05 50 Streptococcus pyrogenes -203 0. 10. 0. 05 25 Bacillus cereus ATCC 10702 0.2 0. 05 1. 56 100 EXAMPLE 1Preparation of 7 (5,6,7,8 tetrahydronaphthyl 1oxyacctylamino)cephalosporanic acid The acid chloride of5,6,7,8-tetrahydronaphthyl-l-oxyacetic acid is prepared by reacting 412mg. (2 mmole) of the acid with excess oxalyl chloride andvacuumevaporating the excess oxalyl chloride. The acid chloride residueis dissolved in 5 ml. of acetone and added dropwise to a stirred,ice-cooled solution prepared by dissolving 548 mg. (2 mmole) of7-aminocephalosporanic acid and 550 mg. (6.6 mmole) of sodiumbicarbonate in 30 ml. of water and 24 m1. of acetone. The reactionmixture is stirred for 2 hours in an ice bath. The acetone isvacuum-evaporated and the residual aqueous solution acidified to pH 2with hydrochloric acid. The solution is extracted three times with 25milliliters each time of ethyl acetate which is then dried overmagnesium sulfate. Evaporation gives 220 mg. of7-(5,6,7,8-tetrahydronaphthyl-1- oxyacetylamino)cephalosporanic acid.The sodium salt is obtained by slurrying this product in 10 ml. of waterand adding dilute sodium hydroxide dropwise to pH 5 to effect solution.Vacuum evaporation to a small volume and addition of a large quantity ofacetone precipitates 204 mg. of sodium7-(5,6,7,B-tetrahydronaphthyl-1-oxyacetylamino)cephalosporanate as awhile solid.

EXAMPLE 2 Preparation of 7-(4-methyl-5,6,7,8tetrahydronaphthyll-oxyacetylamino) cephalosporalnic acid By replacingthe 5,6,7,8-tetrahydronaphthyl-l-oxyacetyl chloride employed in Example1 with an equimolecular quantity of 4 methyl 5,6,'7,8tetrahydronaphthyl-l-oxyacetyl bromide and following substantially thesame procedure described in Example 1, there is obtained the7-(4-rnethyl-5,6,7,8 tetrahydr-onaphthyl-loxyacetylamino)cephalosporanicacid.

EXAMPLE 3 Preparation 0 f 7 (3 -m ethy 1-5 ,6, 7, 7-tetralhydronaph thyl- -oxyacetylamin0 cephalosporanic acid The procedure of Example 1 isrepeated, substituting an equimolecular amount of3-methyl-5,6,7,8-tetrahydronaphthyl-l-oxyacetyl azide for the5,6,7,8-tetrahydronaphtyl-l-oxyacetyl chloride employed in that example.There is thus obtained the 7-(3-methyl-5,6,7,8-tetrahydronaphthyll-oxyacetylamino)cephalosporanic acid.

EXAMPLE 4 Preparation of 7-(1-methy1-5,6,7,8-tetrahydronaphthyl- 2-oxycetylamino)cephalosporanic acid In place of the5,6,7,8-tetrahydronaphthyl-l-oxyacetyl chloride of Example 1, there isemployed an equimolecular quantity of p-nitrophenyl '1-methyl-5,6,7,8-tetrahydronaphthyl-2-oxyacetate whereby the7-(l-methyl-5,6,7,8-tetrahydronaphthyl-2-oxyacetylarnino)cephalosporanicacid is obtained in equally good yield.

EXAMPLE 5 Preparation of 7- (2,4-dichlor0-5,6,7,8-tetrahydronaphth l- 1-0xyacetylamin0) cephalosporanic acid In the manner described in Example1, treatment of 7-arninocephalosporanic acid with 2,4-dichloro-5,'6,7,8-tetrahydronaphthyl-l-oxyacetyl chloride produces the 7 -'(2, 4 dichloro5,6,7,8-tetrahydronaphthyl-l-oxyacetylamino)cephalosporanic acid.

EXAMPLE 6 Preparation of 7-(5,6,7,8-tetrahydronaphthyl- 2-oxycetylamino) cephalosporanic acid The acid chloride of5,6,7,8-tetrahydronaphthyl-Z-oxyacetic acid is prepared by reacting 412mg. (2 mmole) of the acid with excess oxalyl chloride andvacuum-evaporating the excess oxalyl chloride. The acid chloride residueis dissolved in 5 ml. of acetone and added dropwise to a stirred,ice-cooled solution prepared by dissolving 548 mg. (2 mmole) of7-amino)cephalosporanic acid and 500 mg. (6.6 mole) of sodiumbicarbonate in =30 ml. of water and 24 ml. of acetone. The reactionmixture is stirred for 2 hours in an ice bath. The acetone is vacuumevaporated and the residual aqueous solution acidified to pH 2 withhydrochloric acid. The solution is extracted three times with 25milliliters each time of ethyl acetate which is then dried overmagnesium sulfate. Evaporation gives 500 mg. of 7(5,6,7,8-tetrahydronaphthyl-2-oxyacetylamino)cephalosporanic acid. Thesodium salt is obtained by slurrying this product in m1. of water andadding dilute sodium hydroxide dropwise to pH 5 to effect solution.Vacuum evaporation to a small volume and addition of a large quantity ofacetone precipitates 390 mg. of sodium 7 (5,6,7,8 tetrahydronaphthyl2-oxyacetylamino)cephalosporanate as a white solid.

EXAMPLE 7 Preparation of 7 1 -chl0r0-5 ,6, 7,8-te trahydronaphthy Z-OxycetyZamino) cephalosporanic acid By replacing the5,6,7,8-tetrahydronaphthyl-Z-oxyacetyl chloride employed in Example 6with an equimolecular quantity of l-chloro-S,6,7,8-tetrahydronaphthyl-2oxyacetyl bromide and following substantially the same proceduredescribed in Example 6, there is obtained the 7-(1-chloro- 5,6,7,8tetrahydronaphthyl 2-oxyacetylamino)cepha1osporanic acid.

EXAMPLE 8 Preparation of 7-(3-chl0r0-5,6,7,8-tetrahydronaphthyl- Z-oxyketylamino) ceph'alosporanic acid The procedure of Example 6 isrepeated, substituting an equimolecular amount of3-chloro-5,6,7,8-tetrahydronaphthyl-2-oxyacetyl azide for the5,6,7,S-tetrahydronaphthyl- 2-oxyacetyl chloride employed in thatexample. There is thus obtained the7-(3-chloro-5,6,7,8-tetrahydronaphthyl- 2-oxyacetylamino)cephalosporanic acid.

EXAMPLE 9 Preparation of 7-(1,3-dichloro-5,6,7,8-tetrahydronaphthyl-Z-OxyacetyIamino) cephalosporainic acid In the manner described inExample 6, treatment of 7-aminocephalosporanic acid with1,3-dichloro-5,6,7,8- tetrahydronaphthyl-Z-oxyacetyl chloride producesthe 7 (1,3dichloro-5,6,7,8-tetrahydronaphthyl-Z-oxyacetylamino)cephalosporanicacid.

EXAMPLE 10 Preparation of 7-(5,6,7,8-tetrahydronaphthyl-I-oxyacetylamino) 3 (1 pyridylmethyl)-3-cephem-4-carboxylic acid betaine One gram of the sodium salt of7-(5,6,7,8-tetrahydronaphthyl-l-oxyacetylamino)cephalosporanic acid and8 ml. of pyridine was dissolved in 50' ml. of water, which was adjustedto pH 6 with acetic acid, and stored under nitrogen for 3 days at 37 C.The solution was evaporated to dryness and the residue was trituratedwith acetone to give 250 mg. of the product.

EXAMPLE 1'1 Preparation of7-(5,6,7,8-tetrahydronaphthyl-Z-oxyacetylamino) 3 (1pyridylmethyl)-3-cephem-4-carboxylic acid betaine One gram of the sodiumsalt of 7-(5,6,7, 8-tetrahydronaphthyl-2-oxyaeetylamino)cephalosporanicacid and 8 ml. of pyridine was dissolved in 50 ml. of water, which wasadjusted to pH 6 with acetic acid, and stored under nitrogen for 3 daysat 37 C. The solution was evaporated to dryness and the residue wastriturated with acetone to give the product.

EXAMPLE 12 Preparation of 5,6,7,8-tetrahydr0naphthyl-1-0xyacetic acid Amixture of 14.8 g. (0.1 mole) of 5,6,7,8-tetrahydrou-naphthol, 18.9 g.(0.2 mole) of chloroacetic acid, 20.0 g. (0.5 mole) of sodium hydroxide,1.0 g. of potassium iodide and 300 ml. of acetone was refluxed forhours. After evaporation of the acetone the residue was dissolved in oneliter of warm water and acidified to pH 7 with hydrochloric acid. Thesolution was filtered, and the filtrate further acidified to give thecrude 5,6,7,8-tetrahydronaphthyl-l-oxyacetic acid. Reprecipitation fromwater 6 (charcoal treatment) gave 18.5 g. of product, M.P. :1'92- 194 C.

EXAMPLE 13 Preparation of2,4-dichloro-5,6,7,8-tetrahydronaphthyll-oxyacetic acid This acid can besynthesized from the known 2,4-dichl0- ro-5,-6,7,8-tetrahydro-l-naphtholin a manner similar to that described for the preparation of the5,6,7,'8-tetrahydronaphthyl-l-oxyacetic acid of Example 12, except thatwherein R R R R R and R are each selected from the group consisting ofhydrogen, methyl, chloro and bromo; A is selected from the groupconsisting of acetoxy and N-pyridinium; and M is selected from the groupconsisting of hydrogen, pharmaceutically acceptable nontoxic cations andan anionic charge when A is N-pyridinrum.

2. A compound according to Formula (I) of claim 1 where R R and R arehydrogen, A is acetoxy and M is hydrogen.

3. A compound according to Formula (I) of claim 1 wherein R is methyl, Rand R are hydrogen, A is acetoxy and M is hydrogen.

4. A compound according to Formula (I) of claim 1 wherein R and R arehydrogen, R is methyl, A is acetoxy and M is hydrogen.

5. A compound according to Formula (I) of claim 1 wherein R and R arechloro, R is hydrogen, A is acetoxy and M is hydrogen.

6. A compound according to Formula (I) of claim 1 wherein R R and R arehydrogen, A is N-pyridinium and M is an anionic charge.

7. A compound according to Formula (II) of claim 1 wherein R R and R arehydrogen, A is acetoxy and M is hydrogen.

8. A compound according to Formula (II) of claim 1 wherein R and R arechloro, R is hydrogen, A is acetoxy and M is hydrogen.

9. A compound according to Formula (II) of claim 1 wherein R is methyl,R and R are hydrogen, A is acetoxy and M is hydrogen.

10. A compound according to Formula (II) of claim 1 wherein R R and Rare hydrogen, A is N-pyridinium and M is an anionic charge.

No references cited.

NICHOLAS S. RIZZO, Primary Examiner.

1. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF THOSE OF THEFORMULAE: